Nicotiana benthamiana is widely used in plant biology: it is an excellent host for studies of viral replication and gene silencing, and is the preferred platform for transient gene expression via agroinfiltration. N. benthamiana has also become important for commercial-scale production of high value medicinal and industrial proteins. Despite this, genomic resources for N. benthamiana are underrepresented and lag far behind other species in the Solanaceae. KTRDC has initiated a project to increase the genetic and genomic tools available for this species. Using next-generation sequencing technology, we have assembled extensive EST libraries from roots, leaves, flowers and developing seed capsules that will be accessible to the research community via a public BLAST server. We are beginning work on a molecular genetic linkage map of the N. benthamiana genome that will be based on simple sequence repeat (SSR), conserved ortholog sequence (COS), and expressed gene loci. This map will enable evolutionary comparisons to the genomes of important solanaceous crop species such as potato, tomato, pepper and tobacco. Recent molecular genetic investigations have shown that there is very little genetic diversity present in research accessions of N. benthamiana. Therefore, the third part of the KTRDC project will be to introduce genomic, phenotypic and biochemical diversity into this species using fast-neutron (FN) radiation, which is known to cause chromosomal deletions in plants. Our initial target is a minimum of 20,000 irradiated lines, and we expect this effort to require several years to complete. We have defined the FN dose for N. benthamiana, and the first set of irradiated plants (M₀ generation) are growing in the greenhouse. Fertile M₀ plants will be self-pollinated, and phenotypic selection will be undertaken in the M₁ generation. (Reprinted with permission)